The human Descemet's membrane and lens capsule: Protein composition and biomechanical properties

Halfter, Willi; Moes, Suzette; Halfter, Kathrin; Schoenenberger, Monica S.; Monnier, Christophe; Kalita, Joanna; Asgeirsson, Daphne O.; Binggeli, Tatjana; Jenoe, Paul; Scholl, Hendrik P. N.; Henrich, P. B.

doi:10.1016/j.exer.2020.108326

Cited by 19 publications

(19 citation statements)

References 51 publications

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“…The DM is a specialised extracellular matrix underlying the corneal endothelium. Critical for corneal integrity, it is composed of different types of laminin and type IV collagen [ 24 ]. In this study we aim to investigate the DM as a substrate for hESC-RPE cells, thus considering this membrane a potential replacement for the Bruch’s membrane.…”

Section: Discussionmentioning

confidence: 99%

Denuded Descemet’s membrane supports human embryonic stem cell-derived retinal pigment epithelial cell culture

et al. 2023

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Recent clinical studies suggest that retinal pigment epithelial (RPE) cell replacement therapy may preserve vision in retinal degenerative diseases. Scaffold-based methods are being tested in ongoing clinical trials for delivering pluripotent-derived RPE cells to the back of the eye. The aim of this study was to investigate human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells survival and behaviour on a decellularized Descemet’s Membrane (DM), which may be of clinical relevance in retinal transplantation. DMs were isolated from human donor corneas and treated with thermolysin. The DM surface topology and the efficiency of the denudation method were evaluated by atomic force microscope, scanning electron microscopy and histology. hESC-RPE cells were seeded onto the endothelial-side surface of decellularized DM in order to determine the potential of the membrane to support hESC-RPE cell culture, alongside maintaining their viability. Integrity of the hESC-RPE monolayer was assessed by measuring transepithelial resistance. RPE-specific gene expression and growth factors secretion were assessed to confirm maturation and functionality of the cells over the new substrate. Thermolysin treatment did not affect the integrity of the tissue, thus ensuring a reliable method to standardize the preparation of decellularized DM. 24 hours post-seeding, hESC-RPE cell attachment and initial proliferation rate over the denuded DM were higher than hESC-RPE cells cultured on tissue culture inserts. On the new matrix, hESC-RPE cells succeeded in forming an intact monolayer with mature tight junctions. The resulting cell culture showed characteristic RPE cell morphology and proper protein localization. Gene expression analysis and VEGF secretion demonstrate DM provides supportive scaffolding and inductive properties to enhance hESC-RPE cells maturation. Decellularized DM was shown to be capable of sustaining hESC-RPE cells culture, thus confirming to be potentially a suitable candidate for retinal cell therapy.

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Section: Discussionmentioning

confidence: 99%

Denuded Descemet’s membrane supports human embryonic stem cell-derived retinal pigment epithelial cell culture

et al. 2023

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“…Here together with lower viscosity characteristics, human Col IV and recombinant LN521™ were used as additional rheological modi ers in the bioink and to improve the cytocompatibility and functionality of the bioink to support the delicate hPSC-CEnCs. The selection of extracellular matrix proteins was guided by the knowledge that DM is enriched with type IV collagen and laminin α5 [34] and based on our previous results that LN521™ supports the differentiation and culture of hPSC-CEnCs [15]. First, we analyzed the shear-thinning properties with viscosity measurements for the bioink with and without cells and demonstrated that only slight but not signi cant increase was seen in the viscosity of the bioink with cells.…”

Section: Discussionmentioning

confidence: 99%

Bioprinting of human pluripotent stem cell derived corneal endothelial cells with hydrazone crosslinked hyaluronic acid bioink

Grönroos,

Mörö,

Puistola

et al. 2023

Preprint

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Background: Human corneal endothelial cells lack regenerative capacity through cell division in vivo. Thus, in the case of trauma or dystrophy, the only treatment modality currently available is corneal tissue or primary corneal endothelial cell transplantation from cadaveric donor with high global shortage. Our ultimate goal is to use the state-of-the-art 3D-bioprint technology for automated production of human partial and full-thickness corneal tissues using human stem cells and functional bioinks. Here, we explored the possibility to bioprint corneal endothelium using human pluripotent stem cell derived corneal endothelial cells and hydrazone crosslinked hyaluronic acid bioink. Methods: Corneal endothelial cells differentiated from human pluripotent stem cells were bioprinted using optimized hydrazone crosslinked hyaluronic acid based bioink. Before bioprinting, the biocompatibility of the bioink with cells was first analyzed with transplantation on ex vivo denuded porcine corneas and on denuded human Descemet membrane. Then bioprinting was proceeded and the viability of human pluripotent stem cell derived corneal endothelial cells after bioprinting was verified with live/dead stainings. Histological and immunofluorescence stainings with ZO1, Na+/K+-ATPase and CD166 were used to confirm corneal endothelial cell phenotype in all experiments and STEM121 marker was used to identify human cells from the ex vivo porcine corneas. Results: The bioink modified for human pluripotent stem cell derived corneal endothelial cells successfully supported the viability and printability of the cells. After 10 days of ex vivo transplantations, STEM121 positive cells were verified on the Descemet membrane of porcine cornea showing the biocompatibility of the bioink. Furthermore, biocompatibility was confirmed on denuded human Descemet membrane showing corneal endothelial like characteristics. Seven days after bioprinting, the corneal endothelial like cells were viable and showed polygonal morphology with expression and native-like localization of ZO-1, Na+/K+-ATPase and CD166. Nevertheless, mesenchymal-like cells in some parts of the cultures were evident and those cells spread underneath the corneal endothelial-like cell layer. Conclusions: Our results demonstrate that human pluripotent stem cell derived corneal endothelial cells can be bioprinted in covalently crosslinked hyaluronic acid bioink. This approach has potential as a corneal endothelium transplant and furthermore, can be used in the mission of bioprinting the full-thickness human cornea.

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“…[ 28 ] The posterior 60% of the stroma is at least 50% weaker than the anterior stroma, with resultant heightened risks for breaks/ectasia/hydrops within the posterior stroma. Furthermore, nanomechanical measurements have shown that the anterior/epithelial surface of DM is stiffer than its posterior/intracameral surface,[ 29 ] implicating that DM may be more prone to breaks for an equivalent rise in IOP, when compared with PDL. Whether PDL breaks, such as those shown by us, are always associated with Haab’s striae, whether they heal and auto-plug over time, whether their existence portends a poor prognosis for corneal clarity over time, and whether they augment corneal decompensation – the answers to these still elude us and would be worth investigating in a future study.…”

Section: Discussionmentioning

confidence: 99%

Posterior corneal morphological changes in primary congenital glaucoma

Gupta

Mahalingam²,

Singh³

et al. 2022

Indian Journal of Ophthalmology

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Purpose: To compare posterior corneal morphology between older treated and younger untreated children with primary congenital glaucoma (PCG) using anterior segment optical coherence tomography (ASOCT) and intraoperative OCT (iOCT), respectively. Methods: In this comparative study, ASOCT of older PCG children were compared with iOCT of younger untreated PCG patients. Differences between the two groups with respect to posterior corneal morphology were studied. Results: Observed morphological patterns within posterior cornea in older treated (age: 72–300 months) children (87 eyes) included Descemet’s membrane (DM) excrescences (70%), thickened DM (35%), intracameral twin protuberances (92%), and DM detachment (26%). Changes within pre-Descemet’s layer (PDL) (28%) included thickening, breaks, and detachments. Extent of Haab’s striae was associated with thickness of DM/PDL complex ( P = 0.008) when analyzed in the treated group. In contrast, in the untreated group ( n = 53 eyes, age 1–63 months), posterior corneal changes were limited to diffuse hyper-reflectivity of the DM/PDL complex, with absence of DM tears. Conclusion: Posterior cornea thickens and Haab’s striae become more circumscribed in eyes of older treated children compared to untreated PCG eyes, probably reflecting a healing response of posterior cornea over time.

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The human Descemet's membrane and lens capsule: Protein composition and biomechanical properties

Cited by 19 publications

References 51 publications

Denuded Descemet’s membrane supports human embryonic stem cell-derived retinal pigment epithelial cell culture

Denuded Descemet’s membrane supports human embryonic stem cell-derived retinal pigment epithelial cell culture

Bioprinting of human pluripotent stem cell derived corneal endothelial cells with hydrazone crosslinked hyaluronic acid bioink

Posterior corneal morphological changes in primary congenital glaucoma

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